The molecular design of a recombinant antimicrobial peptide CP and its in vitro activity

• Published in: Protein expression and purification 2008, Vol.57 (1), p.95-100
• Main Authors: Niu, Mingfu, Li, Xiang, Wei, Jianchao, Cao, Ruibing, Zhou, Bin, Chen, Puyan
• Format: Article
• Language: English
• Subjects: A synthetic antimicrobial peptide cp gene; Acids - pharmacology; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - pharmacology; Antibacterial activity; Antimicrobial Cationic Peptides - biosynthesis; Antimicrobial Cationic Peptides - chemistry; Antimicrobial Cationic Peptides - genetics; Antimicrobial Cationic Peptides - pharmacology; Bacteria - drug effects; Bacteria - genetics; Cloning, Molecular; Drug Design; Expression; Genes, Bacterial; Genetic Vectors; Gram-Negative Bacteria - drug effects; Gram-Positive Bacteria - drug effects; Hot Temperature; In Vitro Techniques; Microbial Sensitivity Tests; Pichia - genetics; Pichia pastoris; Pichia pastoris X-33; Plasmids; Promoter Regions, Genetic; Protein Precursors - secretion; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - pharmacology; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins - secretion; Stability; Transformation, Genetic
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2007.08.006

Antibacterial peptides from various sources express different antibacterial activity. In order to obtain a high activity antibacterial peptide, the sequences of four antimicrobial peptides—Protegrin-1, 4 kDa Scorpion Defensin, Metalnikowin-2A and Sheep Myeloid Antibacterial Peptide SMAP-29—were exploited to generate a synthetic antimicrobial peptide cp gene, which was then cloned into the expression vector pPICZα-A. The constructed recombinant expression vector pPICZα- cp was transformed into Pichia pastoris X-33, in which the synthetic antimicrobial peptide (CP) could be expressed under the control of the inducible AOX1 promoter and secreted via the α mating factor leader of Saccharomyces cerevisiae. Results showed that recombinant plasmid is highly stable, and In vitro experiments showed that the recombinant antimicrobial peptide CP is heat and acid-stable, and it has high antibacterial activity against several Gram-positive and -negative bacteria. Only 1 μg of the recombinant antimicrobial peptide CP has an antibacterial activity equivalent to 64 U ampicillin. Thus, this recombinant antimicrobial peptide could serve as an attractive candidate for the development of therapeutic antimicrobial drugs.